Method for Producing Triacetin from Tobaccl

ABSTRACT

A method for producing triacetin from one or more plants of genus  Nicotiana  is provided. The triacetin can be derived inter alia from  Nicotiana  species biomass or from seed. In certain embodiments, the triacetin is produced by condensation of glycerin, generated by hydrolysis of acylglycerol-containing starting material derived from tobacco biomass or seed, and acetic acid, also derived from tobacco biomass or seed. The invention also provides articles and compositions including tobacco articles and tobacco compositions that include triacetin produced from one or more plants of genus  Nicotiana.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. patent applicationSer. No. 13/117,655, filed on May 27, 2011, which is incorporated hereinby reference.

FIELD OF THE INVENTION

The present invention relates to products made or derived from tobaccoor, more generally, made or derived from any biomass derived from anyone or more species of genus Nicotiana, or that otherwise incorporatetobacco, and are intended for human consumption. Of particular interestare ingredients or components obtained or derived from plants orportions of plants from the Nicotiana species.

BACKGROUND OF THE INVENTION

Popular smoking articles, such as cigarettes, have a substantiallycylindrical rod shaped structure and include a charge, roll or column ofsmokable material such as shredded tobacco (e.g., in cut filler form)surrounded by a paper wrapper thereby forming a so-called “tobacco rod.”Normally, a cigarette has a cylindrical filter element aligned in anend-to-end relationship with the tobacco rod. Typically, a filterelement comprises plasticized cellulose acetate tow circumscribed by apaper material known as “plug wrap.” Certain cigarettes incorporate afilter element having multiple segments, and one of those segments cancomprise activated charcoal particles. Typically, the filter element isattached to one end of the tobacco rod using a circumscribing wrappingmaterial known as “tipping paper.” It also has become desirable toperforate the tipping material and plug wrap, in order to providedilution of drawn mainstream smoke with ambient air. A cigarette isemployed by a smoker by lighting one end thereof and burning the tobaccorod. The smoker then receives mainstream smoke into his/her mouth bydrawing on the opposite end (e.g., the filter end) of the cigarette.

The tobacco used for cigarette manufacture is typically used in blendedform. For example, certain popular tobacco blends, commonly referred toas “American blends,” comprise mixtures of flue-cured tobacco, burleytobacco, and Oriental tobacco, and in many cases, certain processedtobaccos, such as reconstituted tobacco and processed tobacco stems. Theprecise amount of each type of tobacco within a tobacco blend used forthe manufacture of a particular cigarette brand varies from brand tobrand. However, for many tobacco blends, flue-cured tobacco makes up arelatively large proportion of the blend, while Oriental tobacco makesup a relatively small proportion of the blend. See, for example, TobaccoEncyclopedia, Voges (Ed.) p. 44-45 (1984), Browne, The Design ofCigarettes, 3^(rd) Ed., p. 43 (1990) and Tobacco Production, Chemistryand Technology, Davis et al. (Eds.) p. 346 (1999).

Through the years, various treatment methods and additives have beenproposed for altering the overall character or nature of tobaccomaterials utilized in tobacco products. For example, additives ortreatment processes have been utilized in order to alter the chemistryor sensory properties of the tobacco material, or in the case ofsmokable tobacco materials, to alter the chemistry or sensory propertiesof mainstream smoke generated by smoking articles including the tobaccomaterial. The sensory attributes of cigarette smoke can be enhanced byincorporating flavoring materials into various components of acigarette. Exemplary flavoring additives include menthol and products ofMaillard reactions, such as pyrazines, aminosugars, and Amadoricompounds. See also, Leffingwell et al., Tobacco Flavoring for SmokingProducts, R.J. Reynolds Tobacco Company (1972), which is incorporatedherein by reference. In some cases, treatment processes involving theuse of heat can impart to the processed tobacco a desired color orvisual character, desired sensory properties, or a desired physicalnature or texture. Various processes for preparing flavorful andaromatic compositions for use in tobacco compositions are set forth inU.S. Pat. No. 3,424,171 to Rooker; U.S. Pat. No. 3,476,118 to Luttich;U.S. Pat. No. 4,150,677 to Osborne, Jr. et al.; U.S. Pat. No. 4,986,286to Roberts et al.; U.S. Pat. No. 5,074,319 to White et al.; U.S. Pat.No. 5,099,862 to White et al.; U.S. Pat. No. 5,235,992 to Sensabaugh,Jr.; U.S. Pat. No. 5,301,694 to Raymond et al.; U.S. Pat. No. 6,298,858to Coleman, III et al.; U.S. Pat. No. 6,325,860 to Coleman, III et al.;U.S. Pat. No. 6,428,624 to Coleman, III et al.; U.S. Pat. No. 6,440,223to Dube et al.; U.S. Pat. No. 6,499,489 to Coleman, III; and U.S. Pat.No. 6,591,841 to White et al.; U.S. Pat. Appl. Publication No.2004/0173228 to Coleman, III; and U.S. application Ser. No. 12/191,751to Coleman, III et al., filed Aug. 14, 2008, each of which isincorporated herein by reference. Additionally, examples ofrepresentative components that can be employed as so-called natural tardiluents in tobacco products are set in PCT WO 2007/012980 to Lipowicz,which is incorporated herein by reference.

Tobacco also may be enjoyed in a so-called “smokeless” form.Particularly popular smokeless tobacco products are employed byinserting some form of processed tobacco or tobacco-containingformulation into the mouth of the user. Various types of smokelesstobacco products are set forth in U.S. Pat. No. 1,376,586 to Schwartz;U.S. Pat. No. 3,696,917 to Levi; U.S. Pat. No. 4,513,756 to Pittman etal.; U.S. Pat. No. 4,528,993 to Sensabaugh, Jr. et al.; U.S. Pat. No.4,624,269 to Story et al.; U.S. Pat. No. 4,987,907 to Townsend; U.S.Pat. No. 5,092,352 to Sprinkle, III et al.; and U.S. Pat. No. 5,387,416to White et al.; U.S. Pat. Appl. Pub. Nos. 2005/0244521 to Strickland etal.; 2008/0196730 to Engstrom et al.; and 2009/0293889 to Kumar et al.;PCT WO 04/095959 to Arnarp et al.; PCT WO 05/063060 to Atchley et al.;PCT WO 05/016036 to Bjorkholm; and PCT WO 05/041699 to Quinter et al.,each of which is incorporated herein by reference. See, for example, thetypes of smokeless tobacco formulations, ingredients, and processingmethodologies set forth in U.S. Pat. No. 6,953,040 to Atchley et al. andU.S. Pat. No. 7,032,601 to Atchley et al., each of which is incorporatedherein by reference.

One type of smokeless tobacco product is referred to as “snuff.”Representative types of moist snuff products, commonly referred to as“snus,” have been manufactured in Europe, particularly in Sweden, by orthrough companies such as Swedish Match AB, Fiedler & Lundgren AB,Gustavus AB, Skandinavisk Tobakskompagni A/S, and Rocker Production AB.Snus products available in the U.S.A. have been marketed under thetradenames Camel Snus Frost, Camel Snus Original and Camel Snus Spice byR. J. Reynolds Tobacco Company. See also, for example, Bryzgalov et al.,1N1800 Life Cycle Assessment, Comparative Life Cycle Assessment ofGeneral Loose and Portion Snus (2005). In addition, certain qualitystandards associated with snus manufacture have been assembled as aso-called GothiaTek standard. Representative smokeless tobacco productsalso have been marketed under the tradenames Oliver Twist by House ofOliver Twist A/S; Copenhagen, Skoal, SkoalDry, Rooster, Red Seal, Husky,and Revel by U.S. Smokeless Tobacco Co.; “taboka” by Philip Morris USA;Levi Garrett, Peachy, Taylor's Pride, Kodiak, Hawken Wintergreen,Grizzly, Dental, Kentucky King, and Mammoth Cave by Conwood Company,LLC; and Camel Orbs, Camel Sticks, and Camel Strips by R. J. ReynoldsTobacco Company.

The sensory attributes of smokeless tobacco can also be enhanced byincorporation of certain flavoring materials. See, for example, U.S.Pat. Appl. Pub. Nos. 2002/0162562 to Williams; 2002/0162563 to Williams;2003/0070687 to Atchley et al.; 2004/0020503 to Williams, 2005/0178398to Breslin et al.; 2006/0191548 to Strickland et al.; 2007/0062549 toHolton, Jr. et al.; 2007/0186941 to Holton, Jr. et al.; 2007/0186942 toStrickland et al.; 2008/0029110 to Dube et al.; 2008/0029116 to Robinsonet al.; 2008/0029117 to Mua et al.; 2008/0173317 to Robinson et al.; and2008/0209586 to Neilsen et al., each of which is incorporated herein byreference.

Found as an ingredient in any of a number of tobacco productformulations is triacetin (CAS 102-76-1). Triacetin is also known, forexample, as 1,2,3-propanetriol triacetate; glyceryl triacetate;triacetyl glycerine; or glycerolum triacetas. While it may serve any ofa number of functions in tobacco product formulations, triacetin isknown in the art to serve as a humectant, a plasticizer, or a solvent.In addition, triacetin may function as a fixative or an antifungal. Aliquid at typical room temperature, triacetin is colorless and viscous.

Triacetin is useful as a component of a variety of tobacco products oras an ingredient in the processing of tobacco. See, for example, U.S.Pat. No. 3,166,078 to Parmele et al.; U.S. Pat. No. 4,715,390 to Nicholset al.; U.S. Pat. No. 5,038,802 to White et al.; U.S. Pat. No. 7,237,558to Clark et al.; and U.S. Pat. No. 7,793,665 to Dube et al., each ofwhich is incorporated herein by reference.

As it should be clear from the foregoing that triacetin is useful in theformulation of various tobacco products, it can also be seen that itwould accordingly be desirable to provide a method for producingtriacetin from tobacco, that is, in particular, from Nicotiana species,for use, inter alia, in tobacco compositions utilized in a variety oftobacco products or in the processing of tobacco.

SUMMARY OF THE INVENTION

The present invention provides materials from Nicotiana species (e.g.,tobacco-derived materials) comprising isolated components from plants ofthe Nicotiana species useful for incorporation into tobacco compositionsutilized in a variety of tobacco products, such as smoking articles andsmokeless tobacco products. The invention also provides methods forisolating components from Nicotiana species (e.g., tobacco materials),and methods for processing those components and tobacco materialsincorporating those components. For example, tobacco-derived materialscan be prepared by subjecting at least a portion of a tobacco plant(e.g., leaves, stalks, roots, or stems) to a separation process, whichtypically can include multiple sequential extraction steps, in order toisolate desired components of the tobacco material.

When used in connection with the invention, the term “acylglycerol”denotes any one or more of the following: monoacylglycerol;diacylglycerol; triacylglycerol.

When used in connection with the invention, the term “biomass” denotesone or more portions of a plant, and in particular denotes substantiallythe entirety of the superterranean portion of a plant, optionallyincluding some or all of the subterranean portion of a plant.Accordingly, the term “biomass” may refer to leaf or to seed or to anyother superterranean portion of a plant, or to any combination thereof,optionally including some or all of the subterranean portion of a plant.

When used in connection with the invention, the term “one or more plantsof genus Nicotiana” denotes any one or more plants of the genusNicotiana of family Solanaceae, including, for example, any one or moreof the following: N. alata, N. arentsii, N. excelsior, N. forgetiana, N.glauca, N. glutinosa, N. gossei, N. kawakamii, N. knightiana, N.langsdorffi, N. otophora, N. setchelli, N. sylvestris, N. tomentosa, N.tomentosiformis, N. undulata, and N. x sanderae, N. africana, N.amplexicaulis, N. benavidesii, N. bonariensis, N. debneyi, N.longiflora, N. maritina, N. megalosiphon, N. occidentalis, N.paniculata, N. plumbaginifolia, N. raimondii, N. rosulata, N. rustica,N. simulans, N. stocktonii, N. suaveolens, N. tabacum, N. umbratica, N.velutina, and N. wigandioides, N. acaulis, N. acuminata, N. attenuata,N. benthamiana, N. cavicola, N. clevelandii, N. cordifolia, N.corymbosa, N. fragrans, N. goodspeedii, N. linearis, N. miersii, N.nudicaulis, N. obtusifolia, N. occidentalis subsp. Hersperis, N.pauciflora, N. petunioides, N. quadrivalvis, N. repanda, N.rotundifolia, N. Solanifolia, N. spegazzinii.

When used in connection with the invention, “alkali metal hydroxide”denotes any one or more of the compounds having formula MOH where M isLi, Na, K, Rb, Cs, or Fr.

When used in connection with the invention, “alkaline earth oxide orhydroxide” denotes any one or more of the compounds having formula MO orM(OH)₂ where M is Be, Mg, Ca, Sr, Ba, or Ra.

When used in connection with the invention, “mineral acid” denotes aninorganic acid and accordingly can refer, for example, to any one ormore of the following: sulfuric acid; phosphoric acid; nitric acid;chloric acid; hydrofluoric acid; hydrochloric acid; hydrobromic acid;hydroiodic acid; chromic acid; sulfurous acid; phosphorous acid; nitrousacid; a halogensulfonic acid HSO₃X wherein X is halogen; perchloricacid; perbromic acid; periodic acid; hydrogen sulfide; hypophosphorousacid; tetrafluoroboric acid; hexafluorophosphoric acid.

When used in connection with the invention, “esterase” denotes any oneor more enzymes catalyzing hydrolysis of an ester linkage. For example,“esterase” can refer to any one or more enzymes belonging to EC 3.1(enzymes acting on ester bonds), including, in particular, any one ormore enzymes belonging to EC 3.1.1 (carboxylic ester hydrolases).“Esterase” can accordingly refer, for example, to any one or moreenzymes belonging to any of the following: EC 3.1.1.1(carboxylesterases); EC 3.1.1.3 (triacylglycerol lipases); EC 3.1.1.6(acetylesterases); EC 3.1.1.23 (acylglycerol lipases).

The use of Nicotiana-derived (e.g., tobacco-derived) materials of thepresent invention enables the preparation of tobacco compositions forsmoking articles or smokeless tobacco compositions that are derivedsubstantially or even entirely from Nicotiana materials. For example, atobacco composition can incorporate tobacco or tobacco-derived materialof some form, including isolated components from Nicotiana species, suchthat at least about 80 weight percent, more typically at least about 90weight percent, or even at least about 95 weight percent (on a dryweight basis), of that tobacco composition consists of tobacco-derivedmaterial.

It has been recognized that there is a need to make fuller use ofmaterial or substance from tobacco, and in particular from plants orportions of plants from the Nicotiana species. Readily availablestarting materials or inputs from plants or portions of plants from theNicotiana species, such starting materials or inputs being useful inparticular for inclusion as starting materials or inputs in a processwhereby material or substance from tobacco can be more fully utilized,include inter alia tobacco biomass and tobacco seed. Tobacco biomass caninclude for example the entirety of the substance of a tobacco plantthat has been harvested whole. Tobacco biomass can include for exampleessentially all of the superterranean parts of a tobacco plant andoptionally can include some or all of the subterranean parts of atobacco plant. Tobacco biomass can include for example the solid portionof a tobacco plant that has been harvested whole, or the solid portionof essentially all of superterranean parts of a tobacco plant, and fromwhich so-called “green juice” has been expelled for example through theaction of a screw press. Tobacco biomass can include for example such asolid portion from which at least a portion of the water has beenremoved by drying. A tobacco seed may occupy only a very small volume,such as a fraction of a cubic millimeter. For this reason, it istypically practical to collect a plurality of tobacco seeds when it ischosen to harvest tobacco seed.

Among ways in which fuller use can be made of material or substance fromtobacco, and in particular from plants or portions of plants from theNicotiana species, are various chemical transformations to which plantsor portions of plants from the Nicotiana species can be subjected. Suchchemical transformations may result in outputs or products having one ormore desired or favorable properties. Such outputs or products maythemselves be useful as starting material or inputs for further usefulprocesses. Among chemical transformations to which plants or portions ofplants from the Nicotiana species can be subjected are hydrolyses,including, for example, acid-catalyzed hydrolyses and base-catalyzedhydrolyses. Acid-catalyzed hydrolyses of esters and base-catalyzedhydrolyses of esters are known in the art. An ester which may be subjectto hydrolysis includes an ester found at the sn-1, sn-2 or sn-3 positionof a triacylglycerol, of a diacylglycerol, or of a monoacylglycerol,such as, in particular, a triacylglycerol, diacylglycerol ormonoacylglycerol present in or derived from a tobacco plant or anyportion thereof, including, for example, tobacco biomass or a tobaccoseed.

Various embodiments of hydrolyses of esters, including acid-catalyzedhydrolyses of esters and base-catalyzed hydrolyses of esters, are knownin the art. An acid suitable for an acid-catalyzed hydrolysis of anester, such as an ester in a triacylglycerol, a diacylglycerol, or amonoacylclycerol, may be a mineral acid. A base suitable for abase-catalyzed hydrolysis of an ester, such as an ester in atriacylglycerol, a diacylglycerol, or a monoacylclycerol, may be ahydroxide of a Group 1 alkali metal monovalent cation, such as lithiumhydroxide, sodium hydroxide, or potassium hydroxide.

It is known in the art that the intermediary metabolism of plants,including, inter alia, plants of genus Nicotiana, produces acetate. See,for example, Hoskin et al. (1953), American Journal of Botany, Vol. 40,No. 7, pp. 502 ff. The acetyl moiety is present within the ubiquitousintermediate acetyl coenzyme A, which participates in a variety ofmetabolic reactions. Accordingly, acetate may readily be prepared from aplant source, such as from a plant source from any of the Nicotianaspecies.

Conditions for condensation reactions of polyhydroxy alcohols such asglycerin with acids such as acetic acid are known in the art. See, forexample, Gelosa et al. (2003), Industrial and Engineering ChemicalResearch, Vol. 42, No. 25, pp. 6536 ff.; Ferreira et al. (2009),Catalysis Communications, Vol. 10, No. 5, pp. 481 ff.; Liu et al.(2011), Green Chemistry, Vol. 13, No. 3, pp. 697 ff.

In an aspect, the invention provides a tobacco composition for use in asmoking article or a smokeless tobacco composition comprising a tobaccomaterial and a triacetin-containing component derived from biomass or aseed of the Nicotiana species, wherein the triacetin-containingcomponent comprises triacetin.

In certain embodiments, a glycerin-containing component according to theinvention is formed using techniques adapted for expressing lipids frombiomass or seed, such as high pressure squeezing or cold pressing.Alternatively, a glycerin-containing component according to theinvention is formed by extracting components from biomass or seed usingappropriate extraction techniques and solvents. Exemplary solventsinclude hydrocarbons such as heptane and hexane. Other separationprocesses can be used, such as chromatography, distillation, filtration,recrystallization, solvent-solvent partitioning, and combinationsthereof. A glycerin-containing component formed using an extractionprocess can be either the solvent-soluble portion or the insolubleresidue of biomass or seed material remaining after solvent extraction.A glycerin-containing component formed using a pressing process may beinter alia a lipid-containing portion of biomass or seed expressed frompressed biomass or seed material.

In certain embodiments, an acetate-containing component according to theinvention is formed using distillation techniques adapted for obtainingacetic acid from biomass or one or more anatomical parts of a plant.Alternatively, an acetate-containing component according to theinvention is formed by extracting components from biomass or one or moreanatomical parts of a plant using appropriate extraction techniques andsolvents. Other separation processes can be used, such aschromatography, filtration, recrystallization, solvent-solventpartitioning, and combinations thereof. An acetate-containing componentformed using an extraction process can be either the solvent-solubleportion or the insoluble residue of biomass or other plant materialremaining after solvent extraction.

A glycerin-containing component according to the invention comprisesglycerin, useful for the production of triacetin. Likewise, anacetate-containing component according to the invention comprisesacetate, also useful for the production of triacetin. It willaccordingly be seen that, when used in connection with the invention, an“acetate-containing component” may comprise acetic acid or a salt ofacetic acid.

A triacetin-containing component according to the invention can be usedas such, or in the form of a chemically transformed triacetin-containingcomponent. For example, a chemical transformation of atriacetin-containing component may include acid/base reaction,hydrolysis, thermal treatment, enzymatic treatment, and combinations ofsuch steps.

In a preferred embodiment, a triacetin-containing component according tothe invention is made from a glycerin-containing component derived frombiomass or one or more anatomical parts of tobacco and anacetate-containing component derived from biomass or one or moreanatomical parts of tobacco.

The invention also provides smoking articles and smokeless tobaccocompositions that include a triacetin-containing component as describedherein. For example, a tobacco composition can incorporate atriacetin-containing component within a casing formulation or a topdressing formulation applied to tobacco strip or as a component of areconstituted tobacco material.

The invention, in an aspect, relates to a method for preparing atriacetin-containing component from biomass or one or more anatomicalparts of the Nicotiana species. In a particular preferred embodiment, amethod for preparing a triacetin-containing component according to theinvention comprises (1) isolating an acylglycerol-containing fractionfrom harvested biomass or seed of the Nicotiana species by subjectingthe harvested biomass or seed or a portion thereof to cold pressing,solvent extraction, chromatography, distillation, filtration,recrystallization, solvent-solvent partitioning, or a combinationthereof to form an isolated acylglycerol-containing fraction; (2)chemically transforming the acylglycerol-containing fraction bysubjecting the acylglycerol-containing fraction to hydrolysis, therebyforming a glycerin-containing component; (3) isolating anacetate-containing component from harvested biomass or one or moreanatomical parts of the Nicotiana species by subjecting the harvestedbiomass or one or more anatomical parts or a portion thereof to coldpressing, solvent extraction, chromatography, distillation, filtration,recrystallization, solvent-solvent partitioning, or a combinationthereof to form an isolated acetate-containing component; (4) reactingthe glycerin-containing component with the acetate-containing componentunder conditions favoring esterification, thereby forming atriacetin-containing component. The method can further include the stepof adding a triacetin-containing component according to the invention toa tobacco composition adapted for use in a smoking article or asmokeless tobacco composition.

In connection with the invention it is accordingly found that a chemicaltransformation including an ester-hydrolysis treatment of plants orportions of plants from the Nicotiana species results in formation of acomposition comprising glycerin and from which glycerin can be isolated;that a chemical transformation including a distillation of plants orportions of plants from the Nicotiana species results in formation of acomposition comprising acetate and from which acetate can be isolated;and that an esterification reaction of glycerin so isolated and acetateso isolated results in formation of triacetin.

Accordingly, in one aspect, the invention provides a method of producingtriacetin from tobacco. Such triacetin is suitable for use in, on, oraround a smoking article or a smokeless tobacco composition comprising atobacco material and a component derived from the Nicotiana species,wherein the component is derived from the Nicotiana species.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention now will be described more fully hereinafter. Thisinvention may, however, be embodied in many different forms and shouldnot be construed as limited to the embodiments set forth herein; rather,these embodiments are provided so that this disclosure will be thoroughand complete, and will fully convey the scope of the invention to thoseskilled in the art. As used in this specification and the claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. Reference to “dry weight percent” or“dry weight basis” refers to weight on the basis of dry ingredients(i.e., all ingredients except water).

The selection of the plant from the Nicotiana species can vary; and inparticular, the types of tobacco or tobaccos may vary. Tobaccos that canbe employed include flue-cured or Virginia (e.g., K326), burley,sun-cured (e.g., Indian Kurnool and Oriental tobaccos, includingKaterini, Prelip, Komotini, Xanthi and Yambol tobaccos), Maryland, dark,dark-fired, dark air cured (e.g., Passanda, Cubano, Jatin and Bezukitobaccos), light air cured (e.g., North Wisconsin and Galpao tobaccos),Indian air cured, Red Russian and Rustica tobaccos, as well as variousother rare or specialty tobaccos. Descriptions of various types oftobaccos, growing practices and harvesting practices are set forth inTobacco Production, Chemistry and Technology, Davis et al. (Eds.)(1999), which is incorporated herein by reference. Variousrepresentative types of plants from the Nicotiana species are set forthin Goodspeed, The Genus Nicotiana, (Chonica Botanica) (1954); U.S. Pat.No. 4,660,577 to Sensabaugh, Jr. et al.; U.S. Pat. No. 5,387,416 toWhite et al. and U.S. Pat. No. 7,025,066 to Lawson et al.; U.S. PatentAppl. Pub. Nos. 2006/0037623 to Lawrence, Jr. and 2008/0245377 toMarshall et al.; each of which is incorporated herein by reference. Ofparticular interest are N. alata, N. arentsii, N. excelsior, N.forgetiana, N. glauca, N. glutinosa, N. gossei, N. kawakamii, N.knightiana, N. langsdorffi, N. otophora, N. setchelli, N. sylvestris, N.tomentosa, N. tomentosiformis, N. undulata, and N. x sanderae. Also ofinterest are N. africana, N. amplexicaulis, N. benavidesii, N.bonariensis, N. debneyi, N. longiflora, N. maritina, N. megalosiphon, N.occidentalis, N. paniculata, N. plumbaginifolia, N. raimondii, N.rosulata, N. rustica, N. simulans, N. stocktonii, N. suaveolens, N.tabacum, N. umbratica, N. velutina, and N. wigandioides. Other plantsfrom the Nicotiana species include N. acaulis, N. acuminata, N.attenuata, N. benthamiana, N. cavicola, N. clevelandii, N. cordifolia,N. corymbosa, N. fragrans, N. goodspeedii, N. linearis, N. miersii, N.nudicaulis, N. obtusifolia, N. occidentalis subsp. Hersperis, N.pauciflora, N. petunioides, N. quadrivalvis, N. repanda, N.rotundifolia, N. solanifolia and N. spegazzinii.

Nicotiana species can be derived using genetic-modification orcrossbreeding techniques (e.g., tobacco plants can be geneticallyengineered or crossbred to increase or decrease production of certaincomponents or to otherwise change certain characteristics orattributes). See, for example, the types of genetic modifications ofplants set forth in U.S. Pat. No. 5,539,093 to Fitzmaurice et al.; U.S.Pat. No. 5,668,295 to Wahab et al.; U.S. Pat. No. 5,705,624 toFitzmaurice et al.; U.S. Pat. No. 5,844,119 to Weigl; U.S. Pat. No.6,730,832 to Dominguez et al.; U.S. Pat. No. 7,173,170 to Liu et al.;U.S. Pat. No. 7,208,659 to Colliver et al.; and U.S. Pat. No. 7,230,160to Benning et al.; U.S. Patent Appl. Pub. No. 2006/0236434 to Conklinget al.; and PCT WO 2008/103935 to Nielsen et al.

For the preparation of smokeless and smokable tobacco products, it istypical for harvested plants of the Nicotiana species to be subjected toa curing process. Descriptions of various types of curing processes forvarious types of tobaccos are set forth in Tobacco Production, Chemistryand Technology, Davis et al. (Eds.) (1999). Exemplary techniques andconditions for curing flue-cured tobacco are set forth in Nestor et al.,Beitrage Tabakforsch. Int., 20, 467-475 (2003) and U.S. Pat. No.6,895,974 to Peele, which are incorporated herein by reference. See,also, for example, U.S. Pat. No. 7,650,892 to Groves et al., which isincorporated herein by reference. Representative techniques andconditions for air curing tobacco are set forth in Roton et al.,Beitrage Tabakforsch. Int., 21, 305-320 (2005) and Staaf et al.,Beitrage Tabakforsch. Int., 21, 321-330 (2005), which are incorporatedherein by reference. Certain types of tobaccos can be subjected toalternative types of curing processes, such as fire curing or suncuring. Preferably, harvested tobaccos that are cured are then aged.

At least a portion of the plant of the Nicotiana species (e.g., at leasta portion of the tobacco portion) can be employed in an immature form.That is, the plant, or at least one portion of that plant, can beharvested before reaching a stage normally regarded as ripe or mature.As such, for example, tobacco can be harvested when the tobacco plant isat the point of a sprout, is commencing leaf formation, is commencingseeding, is commencing flowering, or the like.

At least a portion of the plant of the Nicotiana species (e.g., at leasta portion of the tobacco portion) can be employed in a mature form. Thatis, the plant, or at least one portion of that plant, can be harvestedwhen that plant (or plant portion) reaches a point that is traditionallyviewed as being ripe, over-ripe or mature. As such, for example, throughthe use of tobacco harvesting techniques conventionally employed byfarmers, Oriental tobacco plants can be harvested, burley tobacco plantscan be harvested, or Virginia tobacco leaves can be harvested or primedby stalk position. After harvest, the plant of the Nicotiana species, orportion thereof, can be used in a green form (e.g., tobacco can be usedwithout being subjected to any curing process). For example, tobacco ingreen form can be frozen, freeze-dried, subjected to irradiation,yellowed, dried, cooked (e.g., roasted, fried or boiled), or otherwisesubjected to storage or treatment for later use. Such tobacco also canbe subjected to aging conditions.

In accordance with the present invention, a tobacco product incorporatestobacco that is combined with some form of biomass or one or moreanatomical parts obtained from, or derived from, a plant of at least oneNicotiana species. That is, a portion of a tobacco product according tothe invention can be composed of some form of biomass or one or moreanatomical parts of a Nicotiana species, such as parts or pieces ofbiomass or one or more anatomical parts, or processed materialsincorporating processed biomass or one or more anatomical parts orcomponents thereof. At least a portion of the tobacco product can becomposed of components of biomass or one or more anatomical parts, suchas ingredients removed from biomass or one or more anatomical parts(e.g., by extraction, distillation, or other types of processingtechniques). At least a portion of the tobacco product can be composedof components derived from biomass or one or more anatomical parts, suchas components collected after subjecting biomass or one or moreanatomical parts to chemical reaction or after subjecting componentscollected from biomass or one or more anatomical parts to chemicalreaction (e.g., acid/base reaction conditions or enzymatic treatment).

The Nicotiana species can be selected for the type of biomass oranatomical part that it produces. For example, plants can be selected onthe basis that those plants produce relatively abundant biomass or seed,produce biomass or seed that incorporate relatively high levels ofspecific desired components, and the like.

The Nicotiana species of plant can be grown under agronomic conditionsso as to promote development of biomass or one or more anatomical parts.Tobacco plants can be grown in greenhouses, growth chambers, or outdoorsin fields, or grown hydroponically.

According to the invention biomass or one or more anatomical parts areharvested from the Nicotiana species of plant. The manner by whichbiomass or one or more anatomical parts are harvested can vary.Typically, essentially all the biomass or anatomical parts can beharvested, and employed as such.

The time of harvest during the life cycle of the plant can vary. Forexample, biomass or one or more anatomical parts can be harvested whenimmature. Alternatively, biomass or one or more anatomical parts can beharvested after the point that the plant has reached maturity.

The post-harvest processing of biomass or one or more anatomical partscan vary. After harvest, the biomass or one or more anatomical parts, orportion thereof, can be used in the harvested form (e.g., the biomasscan be used without being subjected to any curing and/or aging processsteps). For example, biomass or one or more anatomical parts can be usedwithout being subjected to significant storage, handling or processingconditions. In certain situations, it is preferable that the freshbiomass or one or more anatomical parts be used virtually immediatelyafter harvest. Alternatively, for example, biomass or one or moreanatomical parts can be refrigerated or frozen for later use, freezedried, subjected to irradiation, yellowed, dried, cured (e.g., using airdrying techniques or techniques that employ application of heat), heatedor cooked (e.g., roasted, fried or boiled), or otherwise subjected tostorage or treatment for later use.

Harvested biomass or seed can be physically processed. Biomass or one ormore anatomical parts, or one or more parts thereof, can be furthersubdivided into parts or pieces (e.g., biomass or seed can becomminuted, pulverized, milled or ground into pieces or parts that canbe characterized as granules, particulates or fine powders). Biomass orone or more anatomical parts, or one or more parts thereof, can besubjected to external forces or pressure (e.g., by being pressed orsubjected to roll treatment). When carrying out such processingconditions, biomass or one or more anatomical parts can have a moisturecontent that approximates its natural moisture content (e.g., itsmoisture content immediately upon harvest), a moisture content achievedby adding moisture to the biomass or a moisture content that resultsfrom the drying of the biomass. For example, powdered, pulverized,ground or milled pieces of biomass or one or more anatomical parts canhave moisture contents of less than about 25 weight percent, often lessthan about 20 weight percent, and frequently less than about 15 weightpercent. Parts or pieces of biomass or one or more anatomical parts canbe used as components of tobacco products without further processing, oralternatively the particulate biomass or anatomical part material can beprocessed further prior to incorporation into a tobacco product.

Harvested biomass or one or more anatomical parts, or componentsthereof, can be subjected to other types of processing conditions. Forexample, components of biomass or one or more anatomical parts can beseparated from one another, or otherwise fractionated into chemicalclasses or mixtures of individual compounds. As used herein, an“isolated biomass component,” “isolated component of one or moreanatomical parts,” “biomass isolate,” or “isolate of one or moreanatomical parts” is a compound or complex mixture of compoundsseparated from biomass or one or more anatomical parts of a plant of theNicotiana species. The isolated biomass component or isolated componentof one or more anatomical parts can be a single compound, a homologousmixture of similar compounds (e.g., isomers of a flavorful or aromaticcompound), or a heterologous mixture of dissimilar compounds (e.g., acomplex mixture of various compounds of different types, preferablyhaving desirable sensory attributes).

Examples of the types of components that can be present in a biomassisolate or an isolate of one or more anatomical parts include variousfatty acids and various triglycerides. Exemplary fatty acids includepalmitic acid, linoleic acid, oleic acid, caprylic acid, myristic acid,pentadecanoic acid, palmetoleic acid, heptadecanoic acid, heptadecenoicacid, elaidic acid, gamma-lenolenic acid, arachidic acid, arachidonicacid, 11-eicosenoic acid, 8,11,14-eicosatrieonic acid, 11,14,17-eicosatrienoic acid, 5,8,11,14,17-eicosopentanoic acid,heniecosenoic acid, lignoceric acid, 4,7,10,15,19-decosahexanoic acid,and stearic acid. Exemplary triglycerides include trilinolein,palmito-di-linolein, di-palmito-linolein, tripalmitin, tristearin, andtriolein. Exemplary components of a biomass isolate or an isolate of oneor more anatomical parts also include a variety of other compoundshaving flavor and aroma characteristics such as amino acids and variouspolyphenols.

Typical separation processes can include one or more process steps suchas solvent extraction (e.g., using polar solvents, non-polar organicsolvents, or supercritical fluids), chromatography, distillation,filtration, cold pressing or other pressure-based techniques,recrystallization, and/or solvent-solvent partitioning. Exemplaryextraction and separation solvents or carriers include water, alcohols(e.g., methanol or ethanol), hydrocarbons (e.g., heptane and hexane),diethyl ether methylene chloride and supercritical carbon dioxide.Exemplary techniques useful for extracting components from Nicotianaspecies are described in U.S. Pat. No. 4,144,895 to Fiore; U.S. Pat. No.4,150,677 to Osborne, Jr. et al.; U.S. Pat. No. 4,267,847 to Reid; U.S.Pat. No. 4,289,147 to Wildman et al.; U.S. Pat. No. 4,351,346 to Brummeret al.; U.S. Pat. No. 4,359,059 to Brummer et al.; U.S. Pat. No.4,506,682 to Muller; U.S. Pat. No. 4,589,428 to Keritsis; U.S. Pat. No.4,605,016 to Soga et al.; U.S. Pat. No. 4,716,911 to Poulose et al.;U.S. Pat. No. 4,727,889 to Niven, Jr. et al.; U.S. Pat. No. 4,887,618 toBernasek et al.; U.S. Pat. No. 4,941,484 to Clapp et al.; U.S. Pat. No.4,967,771 to Fagg et al.; U.S. Pat. No. 4,986,286 to Roberts et al.;U.S. Pat. No. 5,005,593 to Fagg et al.; U.S. Pat. No. 5,018,540 toGrubbs et al.; U.S. Pat. No. 5,060,669 to White et al.; U.S. Pat. No.5,065,775 to Fagg; U.S. Pat. No. 5,074,319 to White et al.; U.S. Pat.No. 5,099,862 to White et al.; U.S. Pat. No. U.S. Pat. No. 5,121,757 toWhite et al.; U.S. Pat. No. 5,131,414 to Fagg; U.S. Pat. No. 5,131,415to Munoz et al.; U.S. Pat. No. 5,148,819 to Fagg; U.S. Pat. No.5,197,494 to Kramer; U.S. Pat. No. 5,230,354 to Smith et al.; U.S. Pat.No. 5,234,008 to Fagg; U.S. Pat. No. 5,243,999 to Smith; U.S. Pat. No.5,301,694 to Raymond et al.; U.S. Pat. No. 5,318,050 to Gonzalez-Parraet al.; U.S. Pat. No. 5,343,879 to Teague; U.S. Pat. No. 5,360,022 toNewton; U.S. Pat. No. 5,435,325 to Clapp et al.; U.S. Pat. No. 5,445,169to Brinkley et al.; U.S. Pat. No. 6,131,584 to Lauterbach; U.S. Pat. No.6,298,859 to Kierulff et al.; U.S. Pat. No. 6,772,767 to Mua et al.; andU.S. Pat. No. 7,337,782 to Thompson, all of which are incorporatedherein by reference. See also, the types of separation techniques setforth in Brandt et al., LC-GC Europe, p. 2-5 (March, 2002) and Wellings,A Practical Handbook of Preparative HPLC (2006), which are incorporatedherein by reference. In addition, the biomass or components thereof canbe subjected to the types of treatments set forth in Ishikawa et al.,Chem. Pharm. Bull., 50, 501-507 (2002); Tienpont et al., Anal. Bioanal.Chem., 373, 46-55 (2002); Ochiai, Gerstel Solutions Worldwide, 6, 17-19(2006); Coleman, III, et al., J. Sci. Food and Agric., 84, 1223-1228(2004); Coleman, III et al., J. Sci. Food and Agric., 85, 2645-2654(2005); Pawliszyn, ed., Applications of Solid Phase Microextraction, RSCChromatography Monographs, (Royal Society of Chemistry, UK) (1999);Sahraoui et al., J. Chrom., 1210, 229-233 (2008); and U.S. Pat. No.5,301,694 to Raymond et al., which are incorporated herein by reference.See also, for example, the types of processing techniques set forth inFrega et al., JAOCS, 68, 29-33 (1991); Patel et al., Tob. Res., 24,44-49 (1998); Giannelos et al., Ind. Crops Prod., 16, 1-9 (2002);Mukhtar et al., Chinese J. Chem., 25, 705-708 (2007); Stanisavljevic etal.,Eur. J. Lipid Sci. Technol., 111, 513-518 (2009); which areincorporated herein by reference.

Other methods of forming a biomass isolate or an isolate of one or moreanatomical parts from tobacco can be employed. For example, such amethod can produce a lipid-containing isolate from a tobacco biomass oranatomical part source. Methods of extracting oil components from plantbiomass or one or more anatomical parts are described, for example, inU.S. Pat. No. 4,008,210 to Steele et al.; U.S. Pat. No. 4,009,290 toOkumori et al.; U.S. Pat. No. 4,045,879 to Witte; U.S. Pat. No.4,122,104 to Witte; U.S. Pat. No. 4,298,540 to Youn et al.; U.S. Pat.No. 4,359,417 to Karnofsky et al.; U.S. Pat. No. 4,456,556 to Grimsby;U.S. Pat. No. 4,456,557 to Grimsby; U.S. Pat. No. 4,466,923 toFriedrich; U.S. Pat. No. 4,515,726 to Sullivan; U.S. Pat. No. 4,847,106to Pike et al.; U.S. Pat. No. 5,077,071 to Strop; U.S. Pat. No.5,296,621 to Roos et al.; U.S. Pat. No. 5,397,571 to Roland et al.; U.S.Pat. No. 5,932,095 to Walters et al.; U.S. Pat. No. 6,083,729 to Martinet al.; U.S. Pat. No. 6,225,483 to Franke; U.S. Pat. No. 6,403,126 toWebster et al.; U.S. Pat. No. 6,414,172 to Garces et al.; U.S. Pat. No.6,417,157 to Wadsworth et al.; U.S. Pat. No. 6,495,175 to Rao et al.;U.S. Pat. No. 6,504,085 to Howard; 6,860,998 to Wilde; U.S. Pat. No.7,074,449 to Holley et al.; and U.S. Pat. No. 7,156,981 to Wilde et al.;and U.S. Patent Appl. Pub. Nos. 2002/0121628 to Kapila et al.;2004/0009242 to Krasutsky et al.; 2005/0042347 to Bathurst et al.;2005/0147722 to Fan et al.; and 2006/0111578 to Arhancet et al., all ofwhich are incorporated by reference herein.

Components of biomass or of one or more anatomical parts can besubjected to conditions so as to cause those components (whether as partof the biomass or of the one or more anatomical parts or in the form ofan isolated component) to undergo chemical transformation. For example,a biomass isolate or an isolate of one or more anatomical parts that hasbeen separated from the biomass or one or more anatomical parts can betreated to cause chemical transformation or can be admixed with otheringredients. Such chemical transformation or modification can result inchanges of certain chemical and physical properties of such biomassisolate or isolate of one or more anatomical parts (e.g., sensoryattributes of such an isolate). Exemplary chemical modificationprocesses can be carried out by acid/base reaction, hydrolysis, heating(e.g., a thermal treatment where the isolate is subjected to an elevatedtemperature such as a temperature of at least about 50° C. or at leastabout 75° C. or at least about 90° C.), and enzymatic treatments (e.g.,using hydrolyase, glycosidase, or glucocidase); and as such, componentsof the isolate can undergo esterification, transesterification, isomericconversion, acetal formation, acetal decomposition, and the like.Additionally, various isolated lipid components of the biomass or one ormore anatomical parts can be subjected to hydrogenation in order toalter the degree of saturation of those components, and hence alter thephysical form or behavior of those components.

In one aspect, biomass or one or more anatomical parts can be coldpressed in order to squeeze lipids from the biomass or one or moreanatomical parts, and those lipid components are collected and isolated;or alternatively the biomass or one or more anatomical parts can besubjected to solvent extraction using a solvent (e.g., a polar solventor a non-polar organic solvent), and the resulting extract is collectedand the extracted components are isolated. Then, any of the variousbiomass components or components of one or more anatomical parts may besubjected to enzymatic treatment to form an enzymatically-treatedmaterial. The enzymatically-treated material then is subjected tosolvent extraction to form a biomass isolate or an isolate of one ormore anatomical parts.

In one embodiment, the separating or isolating process comprisesfreezing harvested biomass or one or more anatomical parts or a portionthereof to form a frozen biomass or anatomical part material, processingthe frozen biomass or anatomical part material into a particulate form,subjecting the particulate biomass or anatomical part material to anenzymatic treatment to chemically alter the particulate biomass oranatomical part material, and extracting the particulate biomass oranatomical part material with a solvent to produce a biomass isolate oran isolate of one or more anatomical parts. Exemplary enzymatictreatments include treatment with a glycosidase or a glucosidase.

The biomass or one or more anatomical parts and components or isolatesthereof are useful as components for tobacco compositions, particularlytobacco compositions incorporated into smoking articles or smokelesstobacco products. Addition of such components according to the inventionto a tobacco composition can enhance a tobacco composition in a varietyof ways, depending on the nature of the biomass or seed isolate and thetype of tobacco composition. Exemplary such components can serve toprovide flavor and/or aroma to a tobacco product (e.g., composition thatalters the sensory characteristics of tobacco compositions or smokederived therefrom).

The form of biomass isolate or isolate of one or more anatomical partscan vary. Typically, such isolate is in a solid, liquid, or semi-solidor gel form. Biomass or seed isolate can be used in concrete, absolute,or neat form. Such isolate can have a dry particulate form, a waxy form,or a thick paste form. Liquid forms include isolates contained withinaqueous or organic solvent carriers.

The biomass or one or more anatomical parts, processed biomass or one ormore anatomical parts, and biomass isolates or isolates from one moreanatomical parts can be employed in a variety of forms. Biomass or oneor more anatomical parts, or an isolate of biomass or of one moreanatomical parts, can be employed as a component of processed tobaccos.In one regard, the biomass or one or more anatomical parts, orcomponents thereof, can be employed within a top dressing formulation,or within a casing formulation for application to tobacco strip (e.g.,using the types of manners and methods set forth in U.S. Pat. No.4,819,668 to Shelar, which is incorporated herein by reference).Alternatively, the biomass or one or more anatomical parts, orcomponents thereof, can be employed as an ingredient of a reconstitutedtobacco material (e.g., using the types of tobacco reconstitutionprocesses generally set forth in U.S. Pat. No. 5,143,097 to Sohn; U.S.Pat. No. 5,159,942 to Brinkley et al.; U.S. Pat. No. 5,598,868 to Jakob;U.S. Pat. No. 5,715,844 to Young; U.S. Pat. No. 5,724,998 to Gellatly;and U.S. Pat. No. 6,216,706 to Kumar, which are incorporated herein byreference). The biomass or one or more anatomical parts, or componentsthereof, also can be incorporated into a cigarette filter (e.g., in thefilter plug, plug wrap, or tipping paper) or incorporated into cigarettewrapping paper, preferably on the inside surface, during the cigarettemanufacturing process. An isolate from biomass or from one or moreanatomical parts which isolate has a waxy or smooth texture can be usedas a coating for the surface of a formed smokeless tobacco product. Anisolate having sticky properties can be used as an adhesive (orcomponent of an adhesive) or binding agent within tobacco products. Anisolate having a oily or liquid character can be used as a solvent(e.g., to be used to replace, or act comparable to, a triglyceride typeof solvent; or to replace a glycol type of solvent as a humectant or asa carrier for casing components).

The biomass or one or more anatomical parts, processed biomass or one ormore anatomical parts, and biomass isolates or isolates from one moreanatomical parts can be incorporated into smoking articles. The biomassor one or more anatomical parts, processed biomass or one or moreanatomical parts, and biomass isolates or isolates from one moreanatomical parts can be admixed with other components that are employedin the manufacture of tobacco products. Exemplary types of furtheringredients that can be admixed with the biomass or anatomical partmaterial include flavorants, fillers, binders, pH adjusters, bufferingagents, colorants, disintegration aids, antioxidants, humectants andpreservatives. Representative tobacco blends, non-tobacco components,and representative cigarettes manufactured therefrom, are set forth inU.S. Pat. No. 4,836,224 to Lawson et al.; U.S. Pat. No. 4,924,888 toPerfetti et al.; U.S. Pat. No. 5,056,537 to Brown et al.; U.S. Pat. No.5,220,930 to Gentry; and U.S. Pat. No. 5,360,023 to Blakley et al.; USPat. Appl. Pub. No. 2002/0000235 to Shafer et al.; and PCT WO 02/37990.Those tobacco materials also can be employed for the manufacture ofthose types of cigarettes that are described in U.S. Pat. No. 4,793,365to Sensabaugh; U.S. Pat. No. 4,917,128 to Clearman et al.; U.S. Pat. No.4,947,974 to Brooks et al.; U.S. Pat. No. 4,961,438 to Korte; U.S. Pat.No. 4,920,990 to Lawrence et al.; U.S. Pat. No. 5,033,483 to Clearman etal.; U.S. Pat. No. 5,074,321 to Gentry et al.; U.S. Pat. No. 5,105,835to Drewett et al.; U.S. Pat. No. 5,178,167 to Riggs et al.; U.S. Pat.No. 5,183,062 to Clearman et al.; U.S. Pat. No. 5,211,684 to Shannon etal.; U.S. Pat. No. 5,247,949 to Deevi et al.; U.S. Pat. No. 5,551,451 toRiggs et al.; U.S. Pat. No. 5,285,798 to Banerjee et al.; U.S. Pat. No.5,593,792 to Farrier et al.; U.S. Pat. No. 5,595,577 to Bensalem et al.;U.S. Pat. No. 5,816,263 to Counts et al.; U.S. Pat. No. 5,819,751 toBarnes et al.; U.S. Pat. No. 6,095,153 to Beven et al.; U.S. Pat. No.6,311,694 to Nichols et al.; and U.S. Pat. No. 6,367,481 to Nichols etal.; U.S. Pat. Appl. Pub. No. 2008/0092912 to Robinson et al.; and PCTWO 97/48294 and PCT WO 98/16125. See, also, those types of commerciallymarketed cigarettes described Chemical and Biological Studies on NewCigarette Prototypes that Heat Instead of Burn Tobacco, R. J. ReynoldsTobacco Company Monograph (1988) and Inhalation Toxicology, 12:5, p.1-58 (2000).

The Nicotiana biomass or one or more anatomical parts, processed biomassor one or more anatomical parts, and biomass isolates or isolates fromone more anatomical parts can be incorporated into smokeless tobaccoproducts, such as loose moist snuff, loose dry snuff, chewing tobacco,pelletized tobacco pieces (e.g., having the shapes of pills, tablets,spheres, coins, beads, obloids or beans), extruded or formed tobaccostrips, pieces, rods, cylinders or sticks, finely divided groundpowders, finely divided or milled agglomerates of powdered pieces andcomponents, flake-like pieces, molded processed tobacco pieces, piecesof tobacco-containing gum, rolls of tape-like films, readilywater-dissolvable or water-dispersible films or strips (e.g., U.S. Pat.App. Pub. No. 2006/0198873 to Chan et al.), or capsule-like materialspossessing an outer shell (e.g., a pliable or hard outer shell that canbe clear, colorless, translucent or highly colored in nature) and aninner region possessing tobacco or tobacco flavor (e.g., a Newtoniamfluid or a thixotropic fluid incorporating tobacco of some form).Various types of smokeless tobacco products are set forth in U.S. Pat.No. 1,376,586 to Schwartz; U.S. Pat. No. 3,696,917 to Levi; U.S. Pat.No. 4,513,756 to Pittman et al.; U.S. Pat. No. 4,528,993 to Sensabaugh,Jr. et al.; U.S. Pat. No. 4,624,269 to Story et al.; U.S. Pat. No.4,987,907 to Townsend; U.S. Pat. No. 5,092,352 to Sprinkle, III et al.;and U.S. Pat. No. 5,387,416 to White et al.; U.S. Pat. App. Pub. Nos.2005/0244521 to Strickland et al. and 2008/0196730 to Engstrom et al.;PCT WO 04/095959 to Arnarp et al.; PCT WO 05/063060 to Atchley et al.;PCT WO 05/016036 to Bjorkholm; and PCT WO 05/041699 to Quinter et al.,each of which is incorporated herein by reference. See also, the typesof smokeless tobacco formulations, ingredients, and processingmethodologies set forth in U.S. Pat. No. 6,953,040 to Atchley et al. andU.S. Pat. No. 7,032,601 to Atchley et al.; U.S. Pat. Appl. Pub. Nos.2002/0162562 to Williams; 2002/0162563 to Williams; 2003/0070687 toAtchley et al.; 2004/0020503 to Williams, 2005/0178398 to Breslin etal.; 2006/0191548 to Strickland et al.; 2007/0062549 to Holton, Jr. etal.; 2007/0186941 to Holton, Jr. et al.; 2007/0186942 to Strickland etal.; 2008/0029110 to Dube et al.; 2008/0029116 to Robinson et al.;2008/0029117 to Mua et al.; 2008/0173317 to Robinson et al.; and2008/0209586 to Neilsen et al., each of which is incorporated herein byreference.

The residue of biomass or anatomical part material remaining aftersubjecting biomass or anatomical part material to a separation process(e.g., cold pressing or solvent extraction) and removing some portion ofthe biomass or one or more anatomical parts can also be incorporatedinto a tobacco product, including any of the tobacco products mentionedherein with regard to biomass or one or more anatomical parts, orisolates therefrom. For example, a residue remaining after cold pressingbiomass or one or more anatomical parts and removing lipid componentscan be used as a tobacco composition component (e.g., as part of areconstituted tobacco material), and incorporated into a smoking articleor a smokeless tobacco composition. The insoluble pulp residue remainingafter solvent extraction of a solvent-soluble portion of a biomass oranatomical part material can likewise be used as a component of atobacco composition.

Certain isolates, such as triglyceride-containing isolates of biomass orof one or more anatomical parts, can be used as components of capsulesused in smoking articles or smokeless tobacco compositions. Inparticular, triglyceride-containing isolates can be combined with aflavorant and used as a diluting agent or carrier within the internalpayload of certain breakable capsules. Typically, such a capsuleaccording to the invention has an outer wall and an internal liquid,solid, or gel payload. The payload is released upon rupture of thecapsule wall. Exemplary capsule-containing tobacco products that couldincorporate such isolates are set forth in U.S. Pat. Appl. Pub. Nos.2004/0261807 to Dube et al.; 2005/0066982 to Clark et al.; 2007/0186941to Holton et al.; 2008/0302373 to Stokes et al.; and 2009/0050163 toHartmann et al., each of which is incorporated herein by reference.

Aspects of the present invention are more fully illustrated by thefollowing example, set forth to illustrate certain aspects of thepresent invention and not to be construed as limiting thereof.

In connection with the invention it is found that glycerin is readilyderived through hydrolytic treatment of chemically or physicallyuntransformed or transformed biomass or seed from the Nicotiana species.Acid-catalyzed hydrolysis or base-catalyzed hydrolysis(“saponification”) is suitable for such production of glycerin.Additionally, enzymatic treatment with a lipase or other suitable enzymeis useful for such production.

In an embodiment, seed is collected from mature Nicotiana plants. Theseed is ground into a powder, and the powder is extracted with hexane.Solvent is then evaporated from the hexane fraction, whether with heat,or at ambient temperature, or through the use of an azeotrope, leavingan oleaginous mass. The oleaginous mass is dissolved in a suitableamphipathic solvent to form an intermediate mixture. A volume of eitheran alkali metal hydroxide or a mineral acid is then added to theintermediate mixture, and reaction is allowed to proceed, in whichglycerol and fatty acids are formed. Excess solvent is then evaporated,leaving a product mixture. The product mixture is extracted with anonpolar solvent, in which case the bulk of glycerol product remains inthe residue, or with a polar solvent, in which case the bulk of glycerolproduct is extracted.

In an embodiment, the invention provides a method for producing glycerinfrom tobacco, the method comprising contacting anacylglycerol-containing starting material derived from a tobacco plantwith a composition comprising an alkali metal hydroxide or a mineralacid for a period of time sufficient for at least some of theacylglycerol contained in the starting material to be hydrolyzed,thereby producing fatty acids and glycerin.

In an embodiment, the invention provides an article of manufacturecomprising glycerin derived from plants or portions of plants from theNicotiana species by hydrolytic treatment of the plants or portions ofplants.

According to the invention, a mineral acid for use in hydrolytictreatment of plants or portions of plants from the Nicotiana species maycomprise an industrially acceptable mineral acid and may particularlycomprise a mineral acid selected from sulfuric acid and hydrochloricacid.

According to the invention, an alkali metal hydroxide for use inhydrolytic treatment of plants or portions of plants from the Nicotianaspecies may comprise an industrially acceptable alkali metal hydroxideand may particularly comprise sodium hydroxide or potassium hydroxide.

According to the invention, conditions for contacting at least a portionof one or more plants from the Nicotiana species with a mineral acid oran alkali metal hydroxide may be chosen according to the discretion ofone skilled in the art, such so-called reaction conditions includingsuch variables as time of reaction, temperature pressure, pH, ionicstrength, rate of mixing, agitation, sparging, aeration and so forth.

According to the invention, conditions for isolating acetate from atleast a portion of one or more plants from the Nicotiana species may bechosen according to the discretion of one skilled in the art, includingconditions for distillation, filtration or other separation, and soforth.

According to the invention, conditions for reacting acetic acid isolatedfrom at least a portion of one or more plants from the Nicotiana specieswith glycerol isolated from at least a portion of one or more plantsfrom the Nicotiana species may be chosen according to the discretion ofone skilled in the art, such so-called reaction conditions includingsuch variables as time of reaction, temperature pressure, pH, ionicstrength, rate of mixing, agitation, sparging, aeration and so forth.

According to the invention, any industrially acceptable treatment, or notreatment, of at least a portion of one or more plants from theNicotiana species may be suitable for use of at least a portion of oneor more plants from the Nicotiana species in making a compositionaccording to the invention, in a method according to the invention, ormaking an article of manufacture according to the invention.

Many modifications and other embodiments of the invention will come tomind to one skilled in the art to which this invention pertains havingthe benefit of the teachings presented in the foregoing description.Therefore, it is to be understood that the invention is not to belimited to the specific embodiments disclosed and that modifications andother embodiments are intended to be included within the scope of theappended claims. Although specific terms are employed herein, they areused in a generic and descriptive sense only and not for purposes oflimitation.

What is claimed:
 1. An article of manufacture comprising: (a) triacetinproduced by a method comprising: (i) contacting anacylglycerol-containing first starting material derived from one or moreplants of genus Nicotiana with a composition comprising an alkali metalhydroxide, an alkaline earth oxide or hydroxide, a mineral acid, or anesterase for a period of time sufficient for at least some of theacylglycerol contained in the starting material to be hydrolyzed,thereby producing an intermediate comprising glycerin; (ii) isolatingglycerin from the intermediate comprising glycerin to form an isolatedglycerin reactant; (iii) subjecting an acetate-containing secondstarting material derived from one or more plants of genus Nicotiana todestructive distillation, thereby producing an intermediate comprisingacetic acid; (iv) isolating acetic acid from the intermediate comprisingacetic acid to form an isolated acetic acid reactant; (v) contacting aquantity of the isolated glycerin reactant with a quantity of theisolated acetic acid reactant under conditions favoring esterification,thereby producing triacetin; and (b) biomass from one or more plants ofgenus Nicotiana or reconstituted tobacco.
 2. The article of manufactureof claim 1, wherein the biomass from one or more plants of genusNicotiana comprises tobacco biomass.
 3. The article of manufacture ofclaim 1, wherein the biomass from one or more plants of genus Nicotianacomprises tobacco leaf.
 4. A composition comprising triacetin producedby a method comprising: (a) contacting an acylglycerol-containing firststarting material derived from one or more plants of genus Nicotianawith a composition comprising an alkali metal hydroxide, an alkalineearth oxide or hydroxide, a mineral acid, or an esterase for a period oftime sufficient for at least some of the acylglycerol contained in thestarting material to be hydrolyzed, thereby producing an intermediatecomprising glycerin; (b) isolating glycerin from the intermediatecomprising glycerin to form an isolated glycerin reactant; (c)subjecting an acetate-containing second starting material derived fromone or more plants of genus Nicotiana to destructive distillation,thereby producing an intermediate comprising acetic acid; (d) isolatingacetic acid from the intermediate comprising acetic acid to form anisolated acetic acid reactant; (e) contacting a quantity of the isolatedglycerin reactant with a quantity of the isolated acetic acid reactantunder conditions favoring esterification, thereby producing triacetin.